Genome-wide association study of semen volume, sperm concentration, testis size, and plasma inhibin B levels

Semen quality is affected by environmental factors, endocrine function abnormalities, and genetic factors. A GWAS recently identified ERBB4 at 2q34 as a genetic locus associated with sperm motility. However, GWASs for human semen volume and sperm concentration have not been conducted. In addition, testis size also reportedly correlates with semen quality, and it is important to identify genes that affect testis size. Reproductive hormones also play an important role in spermatogenesis. To date, genetic loci associated with plasma testosterone, sex hormone-binding globulin (SHBG), follicle stimulating hormone (FSH), and luteinizing hormone (LH) levels have been identified using GWASs. However, GWASs have not identified any relevant loci for plasma inhibin B levels. We conducted a two-stage GWAS using 811 Japanese men in a discovery stage followed by a replication stage using an additional 721 Japanese men. The results of the discovery and replication stages were combined into a meta-analysis. After setting a suggestive significance threshold for P values < 5 × 10-6　in the discovery stage, we identified ten regions with SNPs (semen volume: one, sperm concentration: three, testes size: two, and inhibin B: four). We selected only the most significant SNP in each region for replication genotyping. Combined discovery and replication results in the meta-analysis showed that the locus 12q21.31 associated with plasma inhibin B levels (rs11116724) had the most significant association (P = 5.7 × 10-8). The LRRIQ1 and TSPAN19 genes are located in the 12q21.31 region. This study provides new susceptibility variants that contribute to plasma inhibin B levels

CA9 and PRELID2; hypoxia-responsive potential therapeutic targets for pancreatic ductal adenocarcinoma as per bioinformatics analyses

A strong hypoxic environment has been observed in pancreatic ductal adenocarcinoma (PDAC) cells, which contributes to drug resistance, tumor progression, and metastasis. Therefore, we performed bioinformatics analyses to investigate potential targets for the treatment of PDAC. To identify potential genes as effective PDAC treatment targets, we selected all genes whose expression level was related to worse overall survival (OS) in The Cancer Genome Atlas (TCGA) database and selected only the genes that matched with the genes upregulated due to hypoxia in pancreatic cancer cells in the dataset obtained from the Gene Expression Omnibus (GEO) database. Although the extracted 107 hypoxia-responsive genes included the genes that were slightly enriched in angiogenic factors, TCGA data analysis revealed that the expression level of endothelial cell (EC) markers did not affect OS. Finally, we selected CA9 and PRELID2 as potential targets for PDAC treatment and elucidated that a CA9 inhibitor, U-104, suppressed pancreatic cancer cell growth more effectively than 5-fluorouracil (5-FU) and PRELID2 siRNA treatment suppressed the cell growth stronger than CA9 siRNA treatment. Thus, we elucidated that specific inhibition of PRELID2 as well as CA9, extracted via exhaustive bioinformatic analyses of clinical datasets, could be a more effective strategy for PDAC treatment

Utility of robot‐assisted radical cystectomy with intracorporeal urinary diversion for muscle‐invasive bladder cancer

Radical cystectomy remains the gold standard for treatment of muscle‐invasive bladder cancer. Robot‐assisted radical cystectomy has technical advantages over laparoscopic radical cystectomy and has emerged as an alternative to open radical cystectomy. Despite the advancements in robotic surgery, experience with total intracorporeal reconstruction of urinary diversion remains limited. Most surgeons have carried out the hybrid approach of robot‐assisted radical cystectomy and extracorporeal reconstruction of urinary diversion, as intracorporeal reconstruction of urinary diversion remains technically challenging. However, intracorporeal reconstruction of urinary diversion might potentially proffer additional benefits, such as decreased fluid loss, reduction in estimated blood loss and a quicker return of bowel function. The adoption of intracorporeal ileal neobladder reconstruction has hitherto been limited to high‐volume academic institutions. In the present review, we compare the totally intracorporeal robot‐assisted radical cystectomy approach with open radical cystectomy and robot‐assisted radical cystectomy + extracorporeal reconstruction of urinary diversion in muscle‐invasive bladder cancer patients

Memory Immune Responses against Pandemic (H1N1) 2009 Influenza Virus Induced by a Whole Particle Vaccine in Cynomolgus Monkeys Carrying Mafa-A1*052∶02

We made an H1N1 vaccine candidate from a virus library consisting of 144 ( = 16 HA×9 NA) non-pathogenic influenza A viruses and examined its protective effects against a pandemic (2009) H1N1 strain using immunologically naïve cynomolgus macaques to exclude preexisting immunity and to employ a preclinical study since preexisting immunity in humans previously vaccinated or infected with influenza virus might make comparison of vaccine efficacy difficult. Furthermore, macaques carrying a major histocompatibility complex class I molecule, Mafa-A1*052∶02, were used to analyze peptide-specific CD8+ T cell responses. Sera of macaques immunized with an inactivated whole particle formulation without addition of an adjuvant showed higher neutralization titers against the vaccine strain A/Hokkaido/2/1981 (H1N1) than did sera of macaques immunized with a split formulation. Neutralization activities against the pandemic strain A/Narita/1/2009 (H1N1) in sera of macaques immunized twice with the split vaccine reached levels similar to those in sera of macaques immunized once with the whole particle vaccine. After inoculation with the pandemic virus, the virus was detected in nasal samples of unvaccinated macaques for 6 days after infection and for 2.67 days and 5.33 days on average in macaques vaccinated with the whole particle vaccine and the split vaccine, respectively. After the challenge infection, recall neutralizing antibody responses against the pandemic virus and CD8+ T cell responses specific for nucleoprotein peptide NP262-270 bound to Mafa-A1*052∶02 in macaques vaccinated with the whole particle vaccine were observed more promptly or more vigorously than those in macaques vaccinated with the split vaccine. These findings demonstrated that the vaccine derived from our virus library was effective for pandemic virus infection in macaques and that the whole particle vaccine conferred more effective memory and broader cross-reactive immune responses to macaques against pandemic influenza virus infection than did the split vaccine

リステリア感染モデルを用いた細胞内微生物認識分子群の機能解析

Listeria monocytogenes(リステリア)はクロファージの細胞質への侵入および細胞内での増殖が可能な細胞内寄生細菌である。本菌をマクロファージに感染させるとカスパーゼ1の活性化が誘導される。しかし、細胞質に侵入できないリステリア変異株はこの応答を誘導しない。そのため、細胞質内に放出されたリステリアの構成成分または本菌の細胞内挙動が何らかの細胞内受容体に認識されることでカスパーゼ1 活性化が誘導されると考えられる。今回、そのような細胞内受容体の同定を試みたところ、細胞質内のDNAを認識する受容体であるAIM2(absent in melanoma 2)がアダプターASC(apoptosis-associated speck-like protein containing a caspase-activating and recruitment domain)を介してリステリアの認識およびカスパーゼ1活性化の誘導に重要な役割を果たすことを見いだした。Listeria monocytogenes invades the cytoplasm of macrophages and induces the activation of caspase-1 and the subsequent maturation of IL-1β and IL-18. Although apoptosis-associated speck-like protein containing a caspase-activating and recruitment domain (ASC), an adaptor protein of Nod-like receptors, has been shown to play an essential role in inducing this cellular response to L. monocytogenes, the mechanism has not been fully elucidated. In this study, we demonstrated that absent in melanoma 2 (AIM2), a cytosolic DNA receptor, plays an important role in the activation of caspase-1 during L. monocytogenes infection

細菌感染におけるインフラマソームの活性化機序と役割の解析

インフラマソームはカスパーゼ1を活性化する自然免疫機構であり、病原体を含む様々な刺激に応じて形成され、炎症性サイトカインの産生やプログラム細胞死を誘導することで感染防御に関わる。本研究では、肺炎球菌が細胞内DNA受容体であるAIM2に認識されてインフラマソームの形成およびカスパーゼ1活性化を誘導することが明らかになった。また、肺炎球菌の肺炎モデルにおいてインフラマソームが防御的に働くことが示された。さらに、リステリアの致死性全身感染においてはインフラマソームが病態を増悪化させることがわかり、免疫抑制性応答へのインフラマソームの関与が初めて明らかになった。Inflammasomes are innate immune mechanisms that activate caspase-1 in response to various stimuli, including microbial pathogens. Once activated, inflammasomes induce the production of pro-inflammatory cytokines and programmed cell death through caspase-1. Accumulating data have suggested inflammasomes play a protective role against microbial infections. In this study, we examined the mechanism and role of inflammasome formation in infection with two Gram-positive bacteria. We found that Streptococcus pneumoniae is recognized by the cytosolic DNA receptor AIM2, which in turn forms an inflammasome complex to activate caspase-1. It was also found that inflammasomes are critical for host defense against pneumococcal pneumonia. On the other hand, inflammasomes were detrimental to the host in lethal infection with Listeria monocytogenes. Our study revealed that inflammasomes can be upstream of an immunosuppressive response, which may explain the detrimental effect of inflammasomes.研究課題/領域番号:23590504, 研究期間(年度):2011-04-01 - 2015-03-3

リステリア感染におけるToll様受容体を介した自然免疫応答の誘導機序の解析

Toll-like receptors(TLR)は、病原体由来の構造パターン(PAMPs)を認識し、侵入した病原体に対する防御に働くと考えられているが、病原体は通常複数のPAMPsを有しており、細胞内で特殊な動態を示す場合もある。そこで、細胞内寄生細菌であるリステリアの感染モデルを用いて各TLRの役割を解析したところ、複数のTLRを含むMyD88依存的なレセプターが補償的に働くことで宿主防御が強固に維持されていることが明らかになった。研究課題/領域番号:19790319, 研究期間(年度):2007-2008出典：「リステリア感染におけるToll様受容体を介した自然免疫応答の誘導機序の解析」研究成果報告書　課題番号19790319（KAKEN：科学研究費助成事業データベース（国立情報学研究所））（https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-19790319/19790319seika/)を加工して作